U.S. patent number 8,609,726 [Application Number 13/616,613] was granted by the patent office on 2013-12-17 for fatty acid composition for treatment of alzheimer's disease and cognitive dysfunction.
This patent grant is currently assigned to Pronova Biopharma Norge AS. The grantee listed for this patent is Morten Bryhn. Invention is credited to Morten Bryhn.
United States Patent |
8,609,726 |
Bryhn |
December 17, 2013 |
Fatty acid composition for treatment of Alzheimer's disease and
cognitive dysfunction
Abstract
This invention relates to the use of a fatty acid composition
comprising at least (all-Z omega-3)-4,7,10,13,16,19-docosahexaenoic
acid (DHA), or derivatives thereof, and (all-Z
omega-3)-5,8,11,14,17-eicosapentaenoic acid (EPA), or derivatives
thereof for manufacturing of a medicinal product or a food stuff
for the treatment and/or prevention of amyloidos-related diseases,
such as Alzheimer's disease, as well as treatment/prevention of
cognitive dysfunction.
Inventors: |
Bryhn; Morten (Svelvik,
NO) |
Applicant: |
Name |
City |
State |
Country |
Type |
Bryhn; Morten |
Svelvik |
N/A |
NO |
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Assignee: |
Pronova Biopharma Norge AS
(NO)
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Family
ID: |
36697653 |
Appl.
No.: |
13/616,613 |
Filed: |
September 14, 2012 |
Prior Publication Data
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Document
Identifier |
Publication Date |
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US 20130011468 A1 |
Jan 10, 2013 |
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Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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11210115 |
Aug 24, 2005 |
8324276 |
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60645604 |
Jan 24, 2005 |
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Foreign Application Priority Data
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Jan 24, 2005 [SE] |
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0500174-8 |
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Current U.S.
Class: |
514/560 |
Current CPC
Class: |
A61K
31/202 (20130101); A61P 25/00 (20180101); A61K
31/685 (20130101); A61P 25/28 (20180101); A61K
31/22 (20130101) |
Current International
Class: |
A61K
31/20 (20060101) |
Field of
Search: |
;514/560 |
References Cited
[Referenced By]
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FR |
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WO |
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WO |
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WO |
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WO |
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WO |
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Jun 2004 |
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WO |
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WO 2005/070411 |
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Aug 2005 |
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WO |
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WO 2006/077495 |
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Jul 2006 |
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WO |
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Primary Examiner: Zarek; Paul
Attorney, Agent or Firm: Finnegan, Henderson, Farabow,
Garrett & Dunner, LLP
Parent Case Text
This application is a continuation of U.S. patent application Ser.
No. 11/210,115, filed Aug. 24, 2005, now U.S. Pat. No. 8,324,276
which claims the benefit of priority to Swedish Application No.
0500174-8, filed Jan. 24, 2005, and U.S. Provisional Application
No. 60/645,604, filed Jan. 24, 2005, all of which are incorporated
herein by reference.
Claims
The invention claimed is:
1. A method for at least one of maintaining cognitive function and
delaying decline in cognitive function in a subject in need
thereof, comprising administering to the subject an effective
amount of a fatty acid composition comprising (all-Z
omega-3)-4,7,10,13,16,19-docosahexaenoic acid (DHA), and (all-Z
omega-3)-5,8,11,14,17-eicosapentaenoic acid (EPA), with a daily
dosage of the composition ranging from 2 g to 10 g, wherein the
composition is chosen from a medicinal product and a supplement,
and further wherein the composition comprises at least 70%
unsaturated omega-3 fatty acids.
2. A method for at least one of maintaining cognitive function and
delaying decline in cognitive function in a subject in need
thereof, comprising administering to the subject an effective
amount of a fatty acid composition comprising (all-Z
omega-3)-4,7,10,13,16,19-docosahexaenoic acid (DHA), and (all-Z
omega-3)-5,8,11,14,17-eicosapentaenoic acid (EPA), with a daily
dosage of the composition ranging from 2 g to 10 g, wherein the
composition is a supplement, and further wherein the composition
comprises at least 70% unsaturated omega-3 fatty acids.
3. The method according to claim 2, wherein EPA and DHA are present
in the composition in an EPA:DHA ratio ranging from about 1:2 to
2:1.55.
4. The method according to claim 2, wherein the weight ratio of
EPA:DHA in the fatty acid composition is 1:X, where X is equal or
greater than 1.
5. The method according to claim 4, wherein EPA and DHA are present
in the composition in an EPA:DHA ratio ranging from 1:1 to 1:8.
6. The method according to claim 5, wherein the combination of EPA
and DHA is present in the composition in an EPA:DHA ratio ranging
from 1:1 to 1:6.
7. The method according to claim 2, wherein the combination of EPA
and DHA is present in the composition in an EPA:DHA ratio of
2:5.
8. The method according to claim 2, wherein the fatty acids in the
composition are present in at least one form chosen from esterified
form, ethyl ester form, salt form, free acid form, and phospholipid
form, or any combinations thereof.
9. The method according to claim 2, wherein the fatty acid
composition comprises a combination of EPA and DHA in triglyceride
form or ethyl ester form.
10. The method according to claim 2, wherein DHA is obtained from
at least one of vegetable, microbial, and animal origins, or
combinations thereof.
11. The method according to claim 2, wherein the DHA, EPA, or both
are obtained from at least one of vegetable, microbial, and animal
origins, or combinations thereof.
12. The method according to claim 2, wherein the EPA, DHA, or both
are produced from marine oil.
13. The method according to claim 12, wherein at least a part of
the EPA, DHA, or both is produced from fish oil.
14. The method according to claim 2, wherein the fatty acid
composition is produced from marine oil.
15. The method according to claim 2, wherein the fatty acid
composition is administered orally to a human or an animal.
16. The method according to claim 2, wherein the decline in
cognitive function is a mild cognitive dysfunction.
17. The method according to claim 2, wherein the fatty acid
composition is in an oral dosage form chosen from a capsule and a
sachet.
18. The method according to claim 2, wherein the fatty acid
composition is administered in an amount providing a daily dosage
ranging from 2 to 6 g in a human.
Description
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a fatty acid composition for the
use in medicinal products as well as in food stuff or food
complements for treatment and/or prevention of amyloidos-related
diseases, such as Alzheimer's, and/or cognitive dysfunction.
Additionally, the present invention relates to methods for treating
and/or preventing amyloidos-related diseases, such as Alzheimer's,
and/or cognitive dysfunction, which method comprises administering
to a patient a therapeutically effective amount of a fatty acid
composition.
BACKGROUND ART
The biological function of a protein depends on its
three-dimensional structure, which is determined by its amino acid
sequence during the process of protein folding. Normal folding is
needed for successful cell functioning and therefore important in
maintaining health. Several types of diseases have been found where
protein misfolding and conformational change are the main causes of
appearance and progression of the diseases (1).
Misfolding of proteins may lead to formation of so called fibrils.
Proteins or fragment of proteins are converted from their normally
soluble forms to insoluble fibrils or plaques, which accumulate in
a variety of organs. The final forms of these aggregates often have
a well-defined pathological anatomical appearance, known as
amyloid.
Despite the range of proteins involved with their unique and
characteristic native folds, the fibrils of the amyloid in which
they are found in the disease states are extremely similar in their
overall appearance. Proteins known to have the propensity of
fibrillar conformation in humans, called precursor proteins, are
making up a list of 21 exponents (3) and the number is increasing.
Usually the protein in the fibril is made up of a small number of
amino acids on average around 20-60 grouping them in the category
of polypeptides rather than proteins.
Proteins are usually made up of an alfa-helix and a beta-sheet.
Amyloid fibrils, however, usually contain beta-sheet material only
rendering the molecules physical properties different from the
parent protein. While normal proteins are subjected to a continuous
process of degradation by proteolysis, one very important feature
of fibrils is the ability, once formed, to be essentially
indestructible under physiological conditions. The amyloid fibrils
are dominated by hydrogen bonding between the amid and the carbonyl
groups of the main chain, rather than by specific interactions of
the side chains, which determine the structure of normal proteins.
This abnormal bonding induced by the large number of hydrogen bonds
of the beta-sheet that must be disrupted to rescue the polypeptide
chain from the aggregated state, results in a high resistance to
degradation and properly removal from the tissue of deposition.
While in alfa-helices the hydrogen bonds are between side groups
within the same strand, in beta sheets the bonds are between one
strand and another. Since the second beta-strand can come from a
different region of the same protein or from a different molecule,
formation of beta-sheets is usually stabilised by protein
oligomerisation or aggregation. In this manner the misfolded
protein self-associates and become deposited in amyloid aggregates
in diverse organs, inducing tissue damage and organ dysfunction. An
important part of the deposition process is that a critical
concentration of the precursor proteins has to be present before
fibril formation occurs (4). It also seems that as soon as an
amyloid nucleus has been created the process of aggregation and
deposition of amyloid material escalates.
Many of the precursor proteins are not directly prone to fibril
deformation. However, when peptide fragments of the precursor
protein dissociate from the parent molecule such peptides do not
have a stable globular fold to protect them against aggregation.
Folding of proteins is a function of physical properties inherent
from the amino acid sequence of the chain. These so called
non-covalent interactions are weak bonding forces, however, the
large number of individual contacts within a protein adds up to a
large energy factor favouring normal protein folding. The most
important force is the hydrophobic interaction but even hydrogen
bonds mentioned above are extremely important. Examples of even
weaker forces are electrostatic interactions and van der Waals
forces. The number of non-covalent interactions is to some degree a
function of the protein chain length meaning that splicing of a
section of the protein to a peptide will render the peptide with
less stability due to the lower number of non-covalent
interactions. The normal folding forces will be weaker which could
favour the formation of fibrils.
Less known but significantly important for normal folding as well
as maintenance of a stable three-dimensional structure, is protein
acylation by covalent attachment of fatty acids (5). It is well
established that the protein albumin is able to bind several
molecules of fatty acids. Saturated fatty acids such as stearic,
palmitic and myristic acid are the predominant fatty acids that
attach to proteins in eukaryotic cells (6). From studies using
radiolabelled fatty acids we know that each fatty acid labels a
different sub-population of proteins with the fatty acid
interacting with basic amino acids such as lysine, glycine and
arginin. The carboxyl group of the fatty acid forms a salt bridge
or a hydrogen bond with basic amino acid side chains. All sites
have cylindrical hydrophobic channels of varying shape that force
the saturated fatty acids to assume a nearly linear configuration.
However, the binding pockets are large enough to accommodate
unsaturated fatty acids such as oleic acid and arachidonic acid
(7).
Interestingly, established amyloid also contains a certain amount
of fatty acids. By methanol extraction of amyloid derived from
transthyretin about 10% of the dry mass was soluble pointing to the
presence of a lipid fraction (8). Gas-chromatography revealed the
presence of mixtures of saturated fatty acids like those mentioned
above, but also to polyunsaturated fatty acids like palmitoleic
acid, linoleic acid, alfa-linolenic acid and arachidonic acid. This
pattern of fatty acids is typical for a modern Western diet, which
is very much based on saturated fat from dairy products and meat
together with seed derived oils. It is quite clear that fatty acids
have a function in the normal folding of proteins. The reason why
fatty acids are found in amyloid is obscure but interestingly
enough the fatty acids found are congruent with the fats of our
diet. One hypothesis is based on the assumption that some fatty
acids bound to the polypeptide or protein have weaker affinity
rendering the chain less stable and therefore prone to fibrillar
deformation.
Amyloid deposits can be reabsorbed and organ function reversed if
the synthesis of amyloidogenic protein is shut down. There seems to
be a fine balance between the rate at which amyloid is formed and
its clearance. It may therefore be possible to promote the
resorption of amyloid by reducing the concentration of the
amyloidogenic protein to a level below a critical threshold without
necessarily eliminating the precursor (AA). Studies of the
mechanism of conversion from normally soluble precursor proteins
into amyloid fibrils have benefited from the fact that the
transition can be reproduced under laboratory conditions. In vitro
experiments have demonstrated that conversion of native, fully
folded protein into a highly amyloidogenic, partially folded
conformer could be blocked by stabilizing native proteins with a
specific ligand (9). Other experiments using native precursor
proteins such as tau-protein (10) and islet amyloid polypeptide
(IAPP) (11) have shown a stimulating effect of certain fatty acids
on the assembly of fibrils and amyloid. All long-chain fatty acids
tested enhanced assembly to some extent, although greater
stimulation was associated with unsaturated forms. Both articles
concluded that polyunsaturated fatty acids such as arachidonic
acid, oleic acid and linoleic acid but also myristic acid exerted
pronounced effects on fibril and amyloid formation. It seemed
therefore that common unsaturated fatty acids in our diet could
stimulate the formation of fibrils and amyloid and consequently
increase the risk of inducing disabling diseases like Alzheimer's
disease.
SUMMARY OF THE INVENTION
Based on the present invention a number of aspects are presented in
the appended claims. These aspects include, but are not limited
to:
1. Use of a fatty acid composition for the treatment and/or
prevention of amyloidos-related diseases, such as Alzheimer's
disease.
2. Use of a fatty acid composition for the treatment and/or
prevention of cognitive dysfunction.
3. Use of a food stuff or food supplement comprising a fatty acid
composition for the treatment and/or prevention of
amyloidos-related diseases, such as Alzheimer's disease.
4. Use of a food stuff or food supplement comprising a fatty acid
composition for the treatment and/or prevention of cognitive
dysfunction.
5. A method for treatment and/or prevention of amyloidos-related
diseases, such as Alzheimer's disease.
6. A method for treatment and/or prevention of cognitive
dysfunction.
As used herein, the term "Alzheimer's disease" is to be taken to
include both Alzheimer's disease, Alzheimer's dementia and
Alzheimer's sclerosis.
A common feature for the above aspects is the use of a fatty acid
composition comprising at least a combination of (all-Z
omega-3)-4,7,10,13,16,19-docosahexaenoic acid (DHA), or derivatives
thereof and (all-Z omega-3)-5,8,11,14,17-eicosapentaenoic acid
(EPA), or derivatives thereof.
According to a first aspect of the invention, the invention relates
to the use of a fatty acid composition comprising at least a
combination of (all-Z omega-3)-4,7,10,13,16,19-docosahexaenoic acid
(DHA), or derivatives thereof, and (all-Z
omega-3)-5,8,11,14,17-eicosapentaenoic acid (EPA), or derivatives
thereof for the production of a medicinal product for the treatment
and/or prevention of amyloidos-related diseases, such as
Alzheimer's disease.
This first aspect also relates to the use of said fatty acid
composition in treatment of amyloidos-related diseases, such as
Alzheimer's disease, as well as a medicinal product for treatment
of Alzheimer's disease comprising said fatty acid composition.
From research leading to the invention it was surprisingly found
that a fatty acid composition according to the invention prevents
formation of so called fibrils or plaques, and/or reduces deposed
fibrils or plaques, known as amyloid. Moreover, a fatty acid
composition according to the invention predominantly containing DHA
and EPA, or derivatives thereof, seemed to prevent and/or delay
formation of fibrils most effectively, wherein the combination of
DHA and EPA may act as an antagonist.
Moreover, the treatment according to the invention could be
preventive reducing the propensity of fibril formation as well as
therapeutic in situations with established amyloid.
Further, under unfavourable conditions, proteins or fragment of
proteins are converted from their normally soluble forms to
insoluble fibrils or plaques, which accumulate in a variety of
organs including the liver, kidneys, spleen, brain, and internal
secretory glands like the beta cells of the pancreas inducing toxic
effects on cells and tissue. The final forms of these aggregates
often have a well-defined pathological anatomical appearance, known
as amyloid. This is the reason for the use of the term amyloidoses
to describe many of the clinical conditions with which deposition
of amyloid are associated. Thus, as used herein, the term
"amyloidos-related" diseases means clinical conditions or diseases
with which deposition of amyloid, preferably as a consequence of
fibril formation, are associated, such as for instance Alzheimer's
disease.
In a second aspect, the present invention relates to the use of a
fatty acid composition comprising at least a combination of (all-Z
omega-3)-4,7,10,13,16,19-docosahexaenoic acid (DHA), or derivatives
thereof, and (all-Z omega-3)-5,8,11,14,17-eicosapentaenoic acid
(EPA), or derivatives thereof, for the production of a medicinal
product for the treatment and/or prevention of cognitive
dysfunction. This second aspect also relates to the use of said
fatty acid composition in the treatment of cognitive dysfunction,
as well as a medicinal product for treatment of cognitive
dysfunction comprising said fatty acid composition.
It was found that the fatty acid composition of the present
invention was effective in decreasing the rate of memory decline in
patients suffering from cognitive dysfunction, for example in
patients suffering from mild cognitive dysfunction.
In a preferred embodiment of both the above first and second
aspects of the present invention, the invention relates to the use
of a fatty acid composition, wherein the weight ratio of EPA:DHA in
the fatty acid composition is 1:2 to 2:1.55.
In another preferred embodiment of both the above first and second
aspects of the present invention, the invention relates to the use
of a fatty acid composition, wherein the weight ratio of EPA:DHA in
the fatty acid composition is 1:X, where X is equal or greater than
1. Please note that X being one of an integer or non-integer.
The term "amount" herein relates to weight or volume of the fatty
acid composition.
In addition, the invention also includes a fatty acid composition
comprising a combination of one or more DHA and/or EPA derivatives.
Moreover, the desired pharmacological and/or therapeutic effect may
be achieved by the fatty acid composition according to the
invention.
In a preferred embodiment of the invention, EPA and DHA in the
fatty acid composition are present in the composition in an EPA:DHA
ratio from 1:1 to 1:8. In a more preferred embodiment the EPA:DHA
ratio in the fatty acid composition is from about 1:1 to 1:6. In a
further embodiment of the invention, the fatty acid composition is
a product containing DHA or a DHA-derivative in combination with
EPA or an EPA-derivative.
Moreover, in another embodiment, the fatty acids in the composition
according to the invention is presented in at least one of
esterified form, ethyl ester form, salt form, phospholipid form and
free acid form, or any combinations thereof. In a preferred
embodiment, the fatty acid composition is comprised of a
combination of EPA and DHA in triglyceride form.
In another embodiment, at least DHA is obtained from at least one
of vegetable, microbial and animal origins, or combinations
thereof. Moreover, in a further embodiment, wherein the fatty acid
composition comprising at least a combination of DHA and EPA, at
least one of DHA and EPA is obtained from at least one of
vegetable, microbial and animal origins or combinations thereof.
The medicinal product or pharmaceutical product includes therefore
for instance a fatty acid composition comprising at least one of a
DHA-containing microbial oil and a mixture of an DHA-containing oil
from microbial origin and a EPA-containing oil from a marine
origin. Moreover, the fatty acid composition according to the
invention may additionally also comprise at least one of
arachidonic acid (ARA), docosapentaenoic acid, heneicosapentaenoic
acid and octadecatentraenoic or derivatives thereof, or any
combinations thereof. Suitably, at least a part of the EPA and/or
DHA is produced from a marine oil, preferably a fish oil.
Furthermore, in another embodiment of the medicinal product, the
fatty acid composition is produced from a marine oil, such as a
fish oil.
In a specific embodiment of the invention, the fatty acid
composition is comprised of at least a combination of EPA and DHA
in triglyceride form. Moreover, it should be pointed out that the
fatty acid composition is administered to a human or an animal,
preferably orally.
In a specific embodiment of the invention, the medicinal product is
in form of a capsule or contained in a sachet. Preferably, the
capsule is flavoured. More preferably, the capsule is a gelatine
capsule which is flavoured. This embodiment also includes a
capsule, therein both the capsules and the encapsulated fatty acid
composition are flavoured. By flavouring the capsule as mentioned
above, the capsule becomes more attractive to the user. However, a
medicinal product according to the invention may also be produced
for administration though any other route where the active
ingredients may be efficiently absorbed and utilized, e.g.
intravenously, subcutaneously, intramuscularly, intranasally,
rectally, vaginally or topically.
In another embodiment, said fatty acid composition is administered
in an amount providing a daily dosage of 1 g to 15 g of said fatty
acid composition for a human. In a preferred embodiment between 2
and 10 g of said fatty acid composition is administered per day,
and in a more preferred embodiment between 2 and 8 g of said fatty
acid composition. The medicinal product or pharmaceutical
composition or pharmaceutical preparation according to the
invention may also comprise other substances such as an inert
vehicle, or pharmaceutical acceptable adjuvants, carriers,
preservatives etc., which are well known in to those skilled in the
art. However, the medicinal product may also be administered to an
animal, such as a pet or a horse. Moreover, it should be pointed
out that the medicinal product may be at least one of an
amyloid-preventing agent or amyloid-deposit decreasing agent.
In another embodiment of the invention, the invention relates to a
use of a fatty acid composition comprising at least (all-Z
omega-3)-4,7,10,13,16,19-docosahexaenoic acid (DHA), or derivatives
thereof and (all-Z omega-3)-5,8,11,14,17-eicosapentaenoic acid
(EPA), or derivatives thereof, that upon administration to a human
or an animal will prevent the formation of aggregates of protein
fibrils or plaque and/or reduce deposed fibrils, for the production
of a medicinal product for the treatment and/or prevention of
amyloidos-related diseases, such as Alzheimer's disease, and/or
cognitive dysfunction.
According to a third aspect of the invention, the present invention
relates to the use of a fatty acid composition comprising at least
(all-Z omega-3)-4,7,10,13,16,19-docosahexaenoic acid (DHA), or
derivatives thereof, and (all-Z
omega-3)-5,8,11,14,17-eicosapentaenoic acid (EPA), or derivatives
thereof, for the production of a food stuff or food supplement for
the treatment and/or prevention of amyloidos-related diseases, such
as Alzheimer's disease.
From research leading to the invention it was surprisingly found
that a fatty acid composition according to the invention prevents
formation of so called fibrils or plaques, and/or reduces deposed
fibrils or plaques, known as amyloid. A fatty acid composition
containing at least a combination of the two fatty acids DHA and
EPA seemed to prevent and/or delay formation of fibrils most
effectively. Moreover, the use according to above leads to the same
advantages and possibilities as mentioned before. The treatment
according to the invention could be preventive, reducing the
propensity of fibril formation, as well as therapeutic in
situations with established amyloid. The definition concerning
"amyloidos-related" diseases is also included for the patent
positions related to a food stuff or a food supplement according to
the invention.
In a fourth aspect, the present invention also provides the use of
a fatty acid composition comprising at least (all-Z
omega-3)-4,7,10,13,16,19-docosahexaenoic acid (DHA), or derivatives
thereof, and (all-Z omega-3)-5,8,11,14,17-eicosapentaenoic acid
(EPA), or derivatives thereof, for the production of a food stuff
or food supplement for the treatment and/or prevention of cognitive
dysfunction.
It was found that the fatty acid composition of the present
invention was effective in decreasing the rate of memory decline in
patients suffering from cognitive dysfunction, for example in
patients suffering from mild cognitive dysfunction.
One advantage of manufacturing and selling a food stuff for the
treatment and/or prevention of amyloidos-related diseases, such as
Alzheimer's disease, or for treatment and/or prevention of
cognitive dysfunction is that such a food stuff will be more easily
accessible for people. In preventive purpose they preferably buy
the product or supplement in a health store and/or a supermarket,
and they do not need to visit a doctor.
In a preferred embodiment of the third and fourth aspects of the
present invention, the invention relates to the use of a fatty acid
composition, wherein the weight ratio of EPA:DHA in the fatty acid
composition is 1:2 to 2:1.55.
In another preferred embodiment of the third and fourth embodiments
of the present invention, the invention relates to the use of a
fatty acid composition, wherein the weight ratio of EPA:DHA in the
fatty acid composition is 1:X, where X is equal or greater than 1.
Please note that X being one of an integer or non-integer. In
addition, the invention also includes a fatty acid composition
comprising at least a combination of one or more DHA and/or EPA
derivatives. Moreover, the desired pharmacological and/or
therapeutic effect may be achieved by the fatty acid composition
according to the invention.
In another embodiment of the invention, EPA and DHA in the fatty
acid composition are present in the composition of an EPA:DHA ratio
from 1:1 to 1:8. In a more preferred embodiment the EPA:DHA ratio
in the fatty acid composition is from about 1:1 to 1:6. In a
specific embodiment of the invention, the fatty acid composition is
a product of DHA or a DHA-derivative and EPA or an
EPA-derivative.
Moreover, in another embodiment, the fatty acids in the composition
according to the invention is presented in at least one of
esterified form, ethyl ester form, salt form, phospholipid form and
free acid form, or any combinations thereof. In a preferred
embodiment, the fatty acid composition is comprised of a
combination of EPA and DHA in triglyceride form.
Further, in another embodiment, at least DHA is obtained from at
least one of vegetable, microbial and animal origins, or
combinations thereof. In preferred embodiment, wherein the fatty
acid composition comprising at least a combination of DHA and EPA,
at least one of DHA and EPA is obtained from at least one of
vegetable, microbial and animal origins or combinations thereof.
The food stuff or food supplement includes therefore, for instance,
a fatty acid composition comprising at least one of a
DHA-containing microbial oil and a mixture of an DHA-containing oil
from microbial origin and a EPA-containing oil from a marine
origin. Further, the fatty acid composition according to the
invention may additionally also comprise at least one of
arachidonic acid (ARA), docosapentaenoic acid, heneicosapentaenoic
acid and octadecatentraenoic or derivatives thereof, or any
combinations thereof. Suitably, at least a part of the EPA and/or
DHA is produced from a marine oil, preferably a fish oil.
Furthermore, in another embodiment of the food stuff or food
supplement, the fatty acid composition is produced from a marine
oil, such as a fish oil.
In a preferred embodiment of the invention, the fatty acid
composition is comprised of at least a combination of EPA and DHA
in triglyceride form. Moreover, it should be pointed out that the
fatty acid composition is administered to a human or an animal,
preferably orally. However, the food stuff or food supplement
according to the invention may also be produced for administration
though any other route as mentioned before.
In a specific embodiment of the invention, the food stuff or food
supplement is in form of a capsule or contained in a sachet.
Preferably, the capsule is flavoured. More preferably, the capsule
is a gelatine capsule which is flavoured. This embodiment also
includes a capsule, therein both the capsules and the encapsulated
fatty acid composition are flavoured. By flavouring the capsule as
mentioned above, the capsule becomes more attractive to the
user.
In another preferred embodiment, said fatty acid composition is
administered in an amount providing a daily dosage of 1 g to 15 g
of said fatty acid composition for a human. In a more preferred
embodiment between 2 and 10 g of said fatty acid composition is
administered per day, and in a most preferred embodiment between 2
and 8 g of said fatty acid composition. The food stuff or food
supplement according to the invention may also comprise other
substances such as an inert vehicle, or pharmaceutical acceptable
adjuvants, carriers, preservatives etc., which are well known in to
those skilled in the art. Additionally, even the food stuff or food
supplement may be administered to an animal such as a pet or a
horse. Moreover, it should be pointed out that the food stuff or
food supplement may be at least one of an amyloid-preventing agent
or amyloid-deposit decreasing agent.
In a specific embodiment, the present invention relates to use of a
fatty acid composition comprising at least DHA, or derivatives
thereof, and EPA, or derivatives thereof, that upon administration
to a human or an animal will prevent the formation of aggregates of
protein fibrils or plaque and/or reduce deposed fibrils, for the
production of a medicinal product for the treatment and/or
prevention of amyloidos-related diseases, such as Alzheimer's
disease, and/or cognitive dysfunction. Moreover, one advantage of
selling a food stuff for prevention of Alzheimer's disease or a
food stuff for preventing cognitive dysfunction is that the food
stuff may help people not to develop these diseases in the
future.
According to a fifth aspect of the invention, the present invention
relates to a method for the treatment and/or prevention of
amyloidos-related diseases, such as Alzheimer's disease, wherein an
effective amount of a fatty acid composition comprising at least a
combination of (all-Z omega-3)-4,7,10,13,16,19-docosahexaenoic acid
(DHA), or derivatives thereof, and (all-Z
omega-3)-5,8,11,14,17-eicosapentaenoic acid (EPA), or derivatives
thereof, is administered to a human or an animal. Herein, "an
effective amount" also includes a therapeutically or a
pharmaceutically active amount of the fatty acid composition. This
expression relates to a dose of said fatty acid composition that
will lead to the desired pharmacological and/or therapeutic effect.
The desired pharmacological and/or therapeutic effect is, as stated
above, achieved by the fatty acid composition according to the
invention. From research leading to the invention it was
surprisingly found that a fatty acid composition according to the
invention prevents formation of so called fibrils or plaques,
and/or reduces deposed fibrils or plaques, known as amyloid.
In a sixth aspect, the present invention also relates to a method
for the treatment and/or prevention of cognitive dysfunction,
wherein an effective amount of a fatty acid composition comprising
at least a combination of (all-Z
omega-3)-4,7,10,13,16,19-docosahexaenoic acid (DHA), or derivatives
thereof, and (all-Z omega-3)-5,8,11,14,17-eicosapentaenoic acid
(EPA), or derivatives thereof, is administered to a human or an
animal.
It was found that the fatty acid composition of the present
invention was effective in decreasing the rate of memory decline in
patients suffering from cognitive dysfunction, for example in
patients suffering from mild cognitive dysfunction.
In a preferred embodiment of the fifth and sixth aspects of the
present invention, the invention relates to the use of a fatty acid
composition, wherein the weight ratio of EPA:DHA in the fatty acid
composition is 1:2 to 2:1.55. In another preferred embodiment of
the fifth and sixth aspect of the invention, the fatty acid
composition the weight ratio of EPA:DHA in the fatty acid
composition is 1:X, where X is equal or greater than 1. Moreover,
the method leads to the same advantages and possibilities as
mentioned before.
In a preferred method for treatment of Alzheimer's disease and/or
cognitive dysfunction according to the invention, the disease is
caused by deposition of amyloid. In a specific embodiment of the
invention, the method of treating Alzheimer's disease and/or
cognitive dysfunction is related to administering to a human
suffering from the disease an effective disease (Alzheimer's and/or
cognitive dysfunction) alleviating amount of a
amyloid-deposit-decreasing agent. Preferably, the
amyloid-deposit-decreasing agent is a fatty acid composition
comprising at least one of a combination of the fatty acids DHA,
derivatives thereof, and EPA, derivatives thereof.
In another embodiment of the invention, the embodiment relates to
use of a fatty acid composition comprising at least a combination
of DHA, or derivatives thereof, and EPA, or derivatives thereof,
that upon administration to a human or an animal will lead to
prevention of, or reduction of, deposition of amyloid, for the
manufacture of a medicament or a food supplement, for the treatment
and/or prevention of Alzheimer's disease, preferably caused by
amyloidosis.
In a specific embodiment of the invention, the fatty acid
composition of the present invention, for use in a medical product,
food stuff, food supplement or method of treatment comprises at
least 70% unsaturated omega-3 fatty acids wherein the fatty acids
DHA and EPA are present in a weight ratio from about 1:2 to
2:1.
As used herein, amyloidos-related conditions or diseases, include
for example Alzheimer's disease. Such diseases/conditions can be
sporadic, inherited or even infectious, and often occur only late
in life even if inherited forms may appear much earlier. Each
disease is associated with a particular protein and aggregates of
these proteins are thought to be the direct origin of the
pathological conditions associated with the disease. Moreover, the
treatment and/or prevention according to administering the fatty
acid composition of the invention may also include at least one of;
treatment due to reduction of amyloid aggregates, prevention of
misfolding of proteins that may lead to formation of so called
fibrils or plaque, treatment due to decreasing of the production of
A.beta.-protein (amyloid beta protein), and prevention and/or
treatment due to inhibiting or slow down the formation of protein
fibrils, aggregates, or plaque. Moreover, the present invention
also includes prevention of fibril accumulation, or formation, by
administering a fatty acid composition according to the
invention.
In addition, as used herein the term "treatment" means both
treatment having a curving or alleviating purpose and the treatment
of a amyloidos-related disease, such as Alzheimer's, or cognitive
dysfunction can be made either acutely or chronically. By
chronically treatment is meant a treatment witch continues for
weeks or years.
BRIEF DESCRIPTION OF THE DRAWINGS
In the studies and examples below reference is made to the
accompanying drawings. Herein, reference is made to the
accompanying drawings on which:
FIG. 1 shows fibril formation of omega-3 preparations during 30
minutes;
FIG. 2 demonstrates fibril formation between the fatty acid
preparations with the most prominent fibril inducing effect, soy
oil and olive oil, compared to the fibril preventing
DHA-concentrate EPAX 2050 (which comprises approximately 20% EPA
and 50% DHA);
FIG. 3 shows preventive effect against fibril formation up to at
least 150 minutes comparing different omega-3 preparations;
FIG. 4 illustrates effects against fibril formation with a
DHA-concentrate compared to olive oil and soy oil; and
FIG. 5 illustrates results from one of the experiments below,
showing the effect on the MMSE-scoring by orally administering
EPAX.RTM. 2050TG to patients suffering from cognitive
dysfunction.
DESCRIPTION OF PREFERRED EMBODIMENTS
A number of preferred embodiments of the invention were performed
in order to demonstrate that a fatty acid composition comprising a
combination of DHA and EPA is effective on the treatment and/or
prevention of amyloidos-related diseases.
Fibril formation is a consequence of misfolding the precursor
proteins. The reason for this abnormal behaviour of forming normal
three-dimensional structures has not been fully elucidated. As
discussed in the background art, it could seem that common
polyunsaturated fatty acids frequently recommended by dietitians to
prevent cardio-vascular disease and cancer, could in fact increase
the propensity of misfolding precursor proteins, thereby inducing
amyloid deposition. However, the present invention disclose results
showing that another marine long-chain polyunsaturated fatty acid,
namely docosahexaenoic acid (DHA), surprisingly seemed to
indicating a preventive effect on amyloid formation by prolonging
the time to spontaneous fibril formation of IAPP.
The present invention discloses the results of experiments with one
synthetic precursor protein, synthetic IAPP, spontaneously forming
fibrils. This peptide was incubated with free fatty acids of marine
origin comparing the effects on fibril formation with fatty acids
like oleic acid and linoleic acid known to stimulate the formation
of amyloid fibrils. Moreover, the present invention also discloses
results of experiments in a semi-vivo model, where pancreatic
islets from transgenic mice producing human IAPP and spontaneously
forming beta-call amyloid were prepared and incubated with oleic
acid and docosahexaenoic acid.
In a first preferred embodiment, the effects of different omega-3
preparations on fibril formation, were studied.
In a second embodiment, the effects of a fatty acid composition
rich in DHA, EPAX 2050.RTM. (a high omega-3 oil), an oil comprising
at least a combination of DHA and EPA, (K85: approximately 460 mg
EPA and 375 mg DHA), and an olive oil, on deposition of amyloid
fibrils in pancreatic islets, were studied.
EXAMPLES
In Vitro Experiments on Fibril Formation
In the first study, the effects on fibril formation of different
omega-3 preparations were studied. The in vitro fibril formation
experiments were performed in small glass test tubes. A stock of
dissolved fatty acids in ethanol was prepared by dissolving fatty
acids in 100% ethanol at a concentration of 10 mM. After mixing
with the same amount of concentrated NaOH, the final concentration
of ethanol was 3%. The fatty acids or fatty acid combinations in
the table below were tested.
TABLE-US-00001 10 mM Fatty acids Mol. weight contained Linoleic
acid 278 27.8 mg (from soybean oil) Oleic acid 280 28.0 mg (from
olive oil) EPAX DHA concentrate, EPAX2050 301 30.1 mg (.apprxeq.500
mg DHA/g and .apprxeq.200 mg EPA/g) EPAX 4510 318 31.8 mg
(.apprxeq.450 mg EPA/g) EPA 95 302 30.2 mg (.apprxeq.950 mg EPA/g)
K85 314 31.4 mg (.apprxeq.465 mg EPA/g and .apprxeq.375 mg DHA/g)
Control synthIAPP + ethanol
Synthetic precursor protein, synthetic IAPP spontaneously forming
fibrils, was dissolved in dimethylsulfoxide (DMSO) at a
concentration of 10 mg/ml. 25 .mu.M was incubated with each one of
the fatty acids, each one in a concentration of 125 .mu.M, in
distilled water. One .mu.l aliquots of each sample were analysed
after 5, 10, 20, 30, 60, 90, 120, 150, 180, 210, and 240 minutes in
the electron microscope after negative contrasting with 2% uranyl
acetate in 50% ethanol. Formation of fibrils was observed by
electron microscopical analysis and recorded as scores arbitrarily
between 0 and 5 in the 30 minutes experiments and between 1 and 2
in the 240 minutes experiments. FIG. 1 demonstrates fibril
formation of the omega-3 preparations during 30 minutes. The
omega-3 concentrate containing predominantly DHA, EPAX 2050, seemed
to prevent or delay the spontaneously formation of fibrils most
effectively. Moreover, FIG. 2 demonstrates fibril formation between
the fatty acid preparation with the most prominent fibril inducing
effect, olive oil and soy oil, compared to the fibril preventing
DHA-concentrate EPAX 2050.
In addition, fibril induction was also followed for 240 minutes to
quantify the fibril preventive effect of the DHA-concentrate. FIG.
3 demonstrates that preventive effect against fibril formation was
evident up till 150 minutes in the experiment comparing different
omega-3 preparations. Similar prevention was obtained in the
experiment with the DHA concentrate compared to olive oil and soy
oil, see FIG. 4.
Thus, this study shows that treatment with a fatty acid composition
rich in DHA leads to prevention of fibril or plaque formation.
Moreover, it seams that DHA act as an fibril inhibitor. At the same
time the invention also shows preventive effect on fibril formation
of a product comprising at least a combination of DHA and EPA,
wherein preferably the amount of DHA EPA. The results also suggest
a specific preventive effect against fibril formation of an omega-3
product of marine origin comprising EPA and DHA as compared with
soy oil and olive oil.
In Vitro Experiments on Amyloid Formation in Pancreatic Islets
In the second study protective effects on amyloid deposition of an
olive oil, a high DHA omega-3 oil, and a composition comprising at
least DHA and EPA, wherein the amount of EPA.gtoreq.DHA, were
studied.
Transgenic mice carrying the human IAPP gene may be used for
studying deposition of amyloid fibrils in the pancreatic islets
(11). Therefore, single pancreatic islets were isolated and
cultured from transgenic mice. Pancreas were removed under sterile
conditions and placed in Hank's balanced salts and finely minced.
Small pieces of tissue were enzymatically digested by collagenase
for 10 minutes in a 37 degree Celsius water bath. The islets were
individually selected under the microscope. Subsequently the islets
were cultured overnight in 24 well cell cluster containing RPMI
1640 medium supplemented with 10% fetal bovine serum, penicillin
100 U/ml), streptomycin (1, 1 mg/ml) and 22, 0 mM glucose at 37
degrees in humidified air containing 5% carbon monoxide.
500 .mu.M of olive oil, high DHA omega-3 oil (EPAX 2050) and K85
oil and 1% fatty acid-free albumin were added to the wells and the
islets were cultured in RPMI medium during 2 weeks. There were
about 60 islets in each well. Congo red from a stock solution was
added to the wells and the islets were examined using light
microscopy. Amyloid deposition is stained with Congo red while
other cell material does not.
No difference was observed between the different islet groups
regarding survival (Table 1).
TABLE-US-00002 No. of living islets Fatty acid Start End EPAX 2050
60 50 K 85 60 52 Olive oil 65 54
The results of this study shows that 4-5% of the cell islets
incubated in the olive oil solution were stained with Congo red
indicating intracellular amyloid deposition. Surprisingly only 1%
of the cells from islets incubated with high DHA, but even also the
K85 concentrate, were stained with Congo red indicating a
protective effect against amyloid deposition. However, the DHA rich
product, EPAX 2050TG, exhibited stronger effect compared to K85
(EPA.gtoreq.DHA). Additionally, the results of this study confirm
effects supporting prevention and treatment of amyloidos-related
diseases influenced of a fatty:acid composition comprising at least
DHA and EPA, or derivatives thereof.
Prevention of Cognitive Decline
The aim of this study was to evaluate the effects of high dose
omega-3 fatty acid supplementation to patients suffering from
Alzheimer's disease (AD-patients) in preventing cognitive decline
and Alzheimer's disease.
From year 2000 to 2004, 204 AD-patients with a mean age 74.+-.9
(range 54-90) years, under choline esterase inhibitor treatment,
were block randomised to either 4 grams daily of omega-3 fatty
acids in triglyceride form (.about.2.8 g DHA and .about.1.2 g EPA,
EPAX.RTM.2050TG, Pronova Biocare) or placebo (4 g linoleic acid)
for 6 months.
After 6 months, all participants received active treatment for 6
more months. Only patients with Mini Mental State Examination
(MMSE) >15 points and with no other serious illness were
included.
Of the 204 patients participating in the study, 174 patients
(73.+-.9 y, 52% women) fulfilled the protocol and are reported
here. The drop out rate (n=30) did not differ between the two
treatment arms.
Mean MMSE initially in all patients were 23.5.+-.4 points for the
group to initially receive treatment and 27.+-.10 points in the
group to initially receive placebo.
In the whole group, the decline in cognitive function, i.e
MMSE-.about.0.8.+-.2.6 points, did not differ according to
randomisation group at the 6-month follow-up.
However, in the subgroup of patients with mild or very mild
cognitive dysfunction, i.e. MMSE>27 points, there was a
significant reduction in MMSE decline rate in the treatment group
(.DELTA.=.about.0.5.+-.1.5 points, i.e. from 28.4.+-.0.9 to
27.9.+-.1.9 points, n=19) over the first 6 month treatment period
in comparison with the placebo group (.DELTA.=.about.2.6.+-.2.8
points, i.e. from 28.6.+-.1.1 to 26.+-.2.1 points, n=12) (p<0.05
between groups).
A similar arrest in decline rate was observed in the placebo group
(MMSE>27 p) when they received active omega-3 fatty acid
treatment between 6 and 12 months (from 26.+-.2.1 to 25.2.+-.3.1
p). The results for the patient subgroup having initial MMSE >27
is shown in FIG. 5.
These results indicates that high doses of a combination of the
omega-3 fatty acids DHA and EPA, given to patients with mild or
very mild cognitive dysfunction and/or mild or very mild
Alzheimer's disease, delayed the rate of cognitive decline
according to MMSE.
Discussion
In the studies presented above, the synthetic precursor protein
IAPP spontaneously forming fibrils were incubated in vitro with a
series of fatty acids of the omega-3 series as well as omega-6 and
omega-9. The omega-3 concentrate containing DHA and EPA seemed to
prevent or delay the spontaneous formation of fibrils while the
omega-6 (soybean oil) and omega-9 (olive oil) fatty acids seemed to
provoke fibril formation. The last pattern is known from previous
experiments (10, 11). The preventive effect of the combination of
DHA and EPA, however, is an unexpected finding. Other omega-3 fatty
acids did not have the same effect as DHA:EPA even if some
preventive effects were observed compared to the omega-6 and 9
oils. Moreover, in the amyloid in vitro model on islets from
transgenic IAPP producing mice DHA, but even another high
omega-concentrate, K85, induced amyloid deposition to a significant
less extent compared to the omega-9 olive oil.
The present findings indicate a novel therapeutic modality to
diseases caused by amyloid deposition. The treatment could be
preventive reducing the propensity of fibril formation as well as
therapeutic in situations with established amyloid.
Finally, the results support the use of a medicinal product, a
pharmaceutical composition, a food stuff or a food supplement,
comprising a fatty acid composition comprising a combination of DHA
and EPA, for the treatment and/or prevention of amyloidos-related
diseases, such as for instance Alzheimer's disease, as well as for
cognitive dysfunction.
The invention shall not be limited to the shown embodiments and
examples.
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